Comparability and repeatability of methods for estimating the dietary intake of the heterocyclic amine contaminant 2-amino-1-methyl-6-phenylimidazo[4,5b]pyridine (PhIP)

Inconsistent risk estimates for dietary heterocyclic amine (HCA) exposure and cancers may be due to differences in exposure assessment methods and the associated measurement error. We evaluated repeatability and comparability of intake estimates of the HCA 2-amino-1-methyl-6-phenylimidazo[4,5b]pyridine (PhIP) among two food frequency questionnaire (FFQ) collections, three diary collections, and three measurements of urinary PhIP and its metabolites in 36 non-smokers in Baltimore, Maryland, during 2004–2005. Collections spanned ～9 months. Method repeatability was characterised with intraclass correlation coefficients (ICCs). Comparability among methods was assessed with Spearman correlation coefficients. Within-subject variability in PhIP intake was comparably high across all methods (ICCs of 0.20, 0.30, and 0.15 for FFQ, diary, and creatinine-adjusted urinary PhIP, respectively). Mean diary-based PhIP intake and mean urinary PhIP concentration were strongly correlated when restricting the analysis to participants with at least one non-zero diary-based estimate of PhIP intake (n?=?15, r?=?0.75, p?=?0.001), but not in the full study population (n?=?36, r?=?0.18, p?=?0.28). Mean PhIP intake from the FFQ was not associated with that either based on the diary or urinary PhIP separately, but was modestly correlated with a metric that combined the diary- and biomarker-based approaches (r?=?0.30, p?=?0.08). The high within-subject variability will result in significantly attenuated associations if a single measure is used to estimate exposure within an epidemiologic study. Improved HCA assessment tools, such as a combination of methods or validated biomarkers that capture long term exposure, are needed.     

MONITORING POLYCYCLIC AROMATIC COMPOUNDS IN ENVIRONMENTAL AND WORKPLACE SAMPLES USING CONVENTIONAL METHODS AND THE AMES MUTAGENICITY ASSAY OF THEIR NITRATED DERIVATIVES

The development of sensitive methods for monitoring polycyclic aromatic compounds (PACs) has been a central focus of industrial hygiene studies in those industries where workers are exposed to petroleum oils, bitumen fumes or fuel combustion products. The work reported here focuses on one aspect of that effort—workplace monitoring of airborne PAC levels in the hot-mix asphalt paving industry. During the manufacture, transport, and roadway application of hot-mix asphalt (HMA), workers are exposed to low levels of bitumen fumes emanating from the hot product. Over the last twenty-five years, concerns about the health effects of these exposures have prompted numerous studies of the airborne levels of asphalt fumes in the workplace. By and large, these studies have shown that PAC exposures are extremely low—often below the detection limits of standard analytical techniques. For the present study, we have used standard industrial hygiene methods, together with a newer, biologically based assay called the Nitration Assay to measure relative ambient levels of fumes and/or PACs in various paving workplace settings. The latter assay was also used to test bitumen fumes generated in the laboratory by a new “microfuming” technique and to determine specific activities of the 16 PAHs designated by the US EPA as priority pollutants. The Nitration Assay takes advantage of two properties of 3–7-ring PACs: the ease with which they can be chemically nitrated and the high mutagenic potency of the nitrated products in the Ames Salmonella mutagenicity assay. Measurements of fumes and nitratable PAC levels on seven different hot-mix paving jobs showed reasonable correlations between the various methods, as well as patterns of exposure consistent with proximity to fume source.     

AP-PCR Fingerprinting for Detection of Genetic Damage Caused by Polycyclic Aromatic Hydrocarbons

Exposure of organisms to genotoxic chemicals results in the formation of stable, covalently bound adducts between the chemical (or one of its metabolites) and the DNA, these adducts may cause mutations and cytogenetic changes. The primary effects of such exposure (i.e. adduct formation) and subsequent effects on the DNA (cytogenetic damage, mutation) may be monitored using a number of assays of varying sensitivity and specificity. Recent developments in molecular biology offer new possibilities for detecting DNA damage. We examined whether DNA fingerprinting by arbitrarily primed polymerase chain reaction (AP-PCR) can reveal differences in the DNA fingerprints of rats and shore crabs exposed to benzo[a]pyrene in the laboratory and of crabs from control and from polluted areas. The results indicate that differences between control and exposed animals were detectable and that DNA fingerprinting by AP-PCR offers a useful alternative biomarker assay for the detection of the genotoxic effects of environmental pollutants.     

Mercury exposure in female artisanal small-scale gold miners (ASGM) in Mongolia: An analysis of human biomonitoring (HBM) data from 2008

Background

Many poor in developing countries have turned to artisanal small-scale gold mining (ASGM) in an attempt to improve their situation. However, the mercury used to extract gold from ore is discharged in vaporized form into the environment, where it poses a hazard for human health.

Methods

As part of an environmental epidemiological study in Mongolia—to evaluate the burden of environmental mercury contamination—urine, blood and hair samples were collected from residents of areas with or without mercury contamination. A total of 200 blood, urine and hair samples were analyzed for mercury and divided into three subgroups according to mercury content: (1) occupational exposure (high/medium); (2) environmental exposure (low); and (3) no exposure. Internal mercury distributions of the subgroups were compared using the Kruskal-Wallis and Mann-Whitney U-test. The Chi-square test and likelihood ratio proportion were used to compare the findings with threshold limits.

Results

The highest values and greatest differences were seen in the urine samples (p < 0.001, Kruskal-Wallis). The occupational group showing the highest exposure with a median mercury level of 4.36 μg/l (control group: 0.10 μg/l, p < 0.001), 7.18 μg/g creatinine and 12 results above the threshold limit HBM I (Human Biomonitoring I). Even participants from the low-exposure subgroup showed elevated mercury levels (median 2.88 μg/l urine and 2.98 μg/g creatinine, p < 0.001), with 10 individuals above the HBM I threshold limits.

Discussion

The body burden resulting from the use of mercury in artisanal gold mining is high not only in the miners themselves, an increased mercury hazard was also found for inhabitants of mining areas who were not actively involved in mining. Public health support measures are urgently needed to alleviate the situation.     

An Assessment of Fish Assemblage Structure in a Large River

The Penobscot River drains the largest watershed in Maine and once provided spawning and rearing habitats to 11 species of diadromous fishes. The construction of dams blocked migrations of these fishes and likely changed the structure and function of fish assemblages throughout the river. The proposed removal of two main‐stem dams, improved upstream fish passage at a third dam, and construction of a fish bypass on a dam obstructing a major tributary is anticipated to increase passage of and improve habitat connectivity for both diadromous and resident fishes. We captured 61 837 fish of 35 species in the Penobscot River and major tributaries, through 114 km of boat electrofishing. Patterns of fish assemblage structure did not change considerably during our sampling; relatively few species contributed to seasonal and annual variability within the main‐stem river, including smallmouth bass Micropterus dolomieu, white sucker Catostomus commersonii, pumpkinseed Lepomis gibbosus, and golden shiner Notemigonus crysoleucas. However, distinct fish assemblages were present among river sections bounded by dams. Many diadromous species were restricted to tidal waters downriver of the Veazie Dam; Fundulus species were also abundant within the tidal river section. Smallmouth bass and pumpkinseed were most prevalent within the Veazie Dam impoundment and the free‐flowing river section immediately upriver, suggesting the importance of both types of habitat that supports multiple life stages of these species. Further upriver, brown bullhead Ameiurus nebulosus, yellow perch Perca flavescens, chain pickerel Esox niger, and cyprinid species were more prevalent than within any other river section. Our findings describe baseline spatial patterns of fish assemblages in the Penobscot River in relation to dams with which to compare assessments after dam removal occurs. Copyright © 2014 John Wiley & Sons, Ltd.     

How freshwater biomonitoring tools vary sub‐seasonally reflects temporary river flow regimes

Characterizing temporary river ecosystem responses to flow regimes is vital for conserving their biodiversity and the services they provide to society. However, freshwater biomonitoring tools rarely reflect community responses to hydrological variations or flow cessation events, and those available have not been widely tested within temporary rivers. This study examines two invertebrate biomonitoring tools characterizing community responses to different flow‐related properties: the “Drought Effect of Habitat Loss on Invertebrates” (DEHLI) and “Lotic‐invertebrate Index for Flow Evaluation” (LIFE), which, respectively reflect community responses to habitat and hydraulic properties associated with changing flow conditions. Sub‐seasonal (monthly) variations of LIFE and DEHLI were explored within two groundwater‐fed intermittent rivers, one dries sporadically (a flashy, karstic hydrology—River Lathkill) and the other dries seasonally (a highly buffered flow regime—South Winterbourne). Biomonitoring tools were highly sensitive to channel drying and also responded to reduced discharges in permanently flowing reaches. Biomonitoring tools captured ecological recovery patterns in the Lathkill following a supra‐seasonal drought. Some unexpected results were observed in the South Winterbourne where LIFE and DEHLI indicated relatively high‐flow conditions despite low discharges occurring during some summer months. This probably reflected macrophyte encroachment, which benefitted certain invertebrates (e.g., marginal‐dwelling taxa) and highlights the importance of considering instream habitat conditions when interpreting flow regime influences on biomonitoring tools. Although LIFE and DEHLI were positively correlated, the latter responded more clearly to drying events, highlighting that communities respond strongly to the disconnection of instream habitats as flows recede. The results highlighted short‐term ecological responses to hydrological variations and the value in adopting sub‐seasonal sampling strategies within temporary rivers. Findings from this study indicate the importance of establishing flow response guilds which group taxa that respond comparably to flow cessation events. Such information could be adopted within biomonitoring practices to better characterize temporary river ecosystem responses to hydrological variations.     

Biomonitoring of biosurfactant production by green fluorescent protein‐marked Bacillus subtilis W1012

BACKGROUND: Biosurfactant production was investigated using two strains of Bacillus subtilis, one being a reference strain (B. subtilis 1012) and the other a recombinant of this (B. subtilis W1012) made able to produce the green fluorescent protein (GFP). RESULTS: Batch cultivations carried out at different initial levels of glucose (G₀) in the presence of 10 g L^?1 casein demonstrated that the reference strain was able to release higher levels of biosurfactants in the medium at 5.0≤G₀≤10 g L^?1 (B_max = 104–110 mg L^?1). The recombinant strain exhibited slightly lower levels of biosurfactants (B_max = 90–104 mg L^?1) but only at higher glucose concentrations (G₀ ≥ 20 g L^?1). Under these nutritional conditions, the fluorescence intensity linked to the production of GFP was shown to be associated with the cell concentration even after achievement of the stationary phase. CONCLUSION: The ability of the genetically‐modified strain to simultaneously overproduce biosurfactant and GFP even at low biomass concentration makes it an interesting candidate for use as a biological indicator to monitor indirectly the biosurfactant production in bioremediation treatments. Copyright © 2008 Society of Chemical Industry     

Availability and comparability of human biomonitoring data across Europe: A case-study on blood-lead levels

Recently, it has become clear that the complexity of environmental health issues requires an approach that takes into account the complexities, interdependencies and uncertainties of the real world. An urgent issue that has surfaced is the need for accurate tools to better describe exposure characterization to environmental chemicals. By including human biomonitoring (HBM) data, a greater precision in exposure and associated risk estimates and more accurate dose-response relationships may be achieved. A restricting issue still is the availability of reliable and comparable HBM data. The aim of the current study was twofold: (1) to find out whether it is practically feasible to collect raw, individual HBM data across Europe; and (2) to evaluate the comparability and use of these HBM data for environmental health impact assessment at a European scale. Blood-lead (B-Pb) was selected as the chemical of choice because of its long history as an environmental pollutant in HBM programs and its known public health relevance. Through literature search and identification of HBM experts across Europe, HBM programs that measured B-Pb were identified and asked to share individual data on age, gender and B-Pb levels. Following this request, more than 20,000 individual data points from 8 European countries were collected. Analysing these data made clear that it is difficult to use disparate data collections because of the inherent variability with respect to the gender and age of participants and calendar-years sampled. When these confounders were taken however, there was no additional variability in B-Pb distributions among different countries. It was concluded that while it is possible to collect HBM data from different sources across Europe, the need to get data from comparable (sub-)populations is essential for appropriate use and interpretation of HBM data for environmental health impact assessment.     

THE SIGNIFICANCE OF POLYCYCLIC AROMATIC HYDROCARBONS AS ENVIRONMENTAL CARCINOGENS. 35 YEARS RESEARCH ON PAH—A RETROSPECTIVE

In vivo studies with laboratory animals as well as in vitro studies with bacteria and mammalian cell cultures have demonstrated that the mutagenic and/or carcinogenic properties of numerous PAHs require metabolic transformation. Metabolism of PAHs has been explored in vitro using cellular microsomal fractions, mammalian cell cultures and later genetically engineered cells expressing cytochromes P450 from several species including humans.

Balancing the carcinogenic potential of some environmental matrices (vehicle exhaust, condensate of hard coal combustion effluents, cigarette smoke condensate, used motor oil) after separation into sub-fractions evidenced that the carcinogenic effect may be attributed almost exclusively to PAH. Mixtures of well-known carcinogenic PAH in concentrations as present in these matrices, however, did not explain the total biological effect. Thus, it had been speculated that either very potent unknown carcinogens are still hidden in the PAH fraction, or that synergistic effects (enzyme induction) play a significant role.

In parallel to these carcinogenicity studies, the metabolism of various PAHs has been investigated in rat liver microsomes from untreated animals as well as from animals pre-treated with inducers of cytochrome P450. It was found that even non-carcinogenic PAHs possess a significant inducing potential. Moreover, in several mammals a highly species-specific metabolism of PAH could be observed allowing a critical view to the extrapolation from animal experiments to the human situation. This was further confirmed by experiments with mammalian cell cultures including human ones as well as by metabolic studies with genetically engineered Chinese hamster V79 cells singularly expressing various cytochrome P450 enzymes from a number of different species (human, rat, mouse, fish). With these cell lines metabolic studies were carried out with a larger number of PAHs as substrates including phenanthrene, pyrene, chrysene, benzo[a]anthracene, benzo[c]phenanthrene, benzo[a]pyrene, dibenzo[a,l]pyrene, and benzo[c]chrysene.

Based on the metabolism results, analytical methods have been developed to determine urinary biomarkers of human PAH exposure. Human biomonitoring studies have been performed with different occupationally exposed individuals as well as within smokers and non-smokers of the general population. Endogenous PAH exposure levels and changes in the urinary excreted metabolic profile depending on exposure level have been determined.     

Assessment of Exposure to PACs in Asphalt Workers: Measurement of Urinary PACs and their Metabolites with an ELISA Kit

An enzyme-linked immunosorbent assay (ELISA) kit made for determination of polycyclic aromatic compounds (PACs) in water was adapted for measuring PACs and their metabolites in urine. This method was then applied to a pilot asphalt worker PAC exposure study. Currently, liquid-liquid extraction with gas chromatography/isotope dilution high-resolution mass spectrometry (GC/HRMS) is the preferred method to determine urinary PAC metabolites. Although sensitive and specific, GC/HRMS is time consuming and costly.

The ELISA method had a range from 14–720 ng/ml 1-hydroxypyrene equivalents with a lower limit of detection (LOD) of 14 ng/ml urine. ELISA and GC/HRMS PAC metabolite measurements had a statistically significant correlation and the PAC ELISA results were indicative of potential asphalt exposure. PAC ELISA is promising as a more rapid and less costly routine method for determining worker exposure to PACs in asphalt emissions.